Search

Tumor necrosis factor-alpha-converting enzyme (TACE) is a metalloprotease which can shed several cytokines from the cell membrane, including receptor activator of NF-kappaB ligand (RANKL). This study aimed to investigate the hypothesis that TACE would be elevated in the gingival crevicular fluid (GCF) of persons with periodontitis. Total TACE amounts in GCF were higher in persons with chronic and aggressive periodontitis than in those with gingivitis or in healthy persons. TACE concentrations in GCF were higher in persons with chronic and aggressive periodontitis than in those with gingivitis, although not significantly higher than in healthy persons. Persons with chronic periodontitis receiving immunosuppressive treatment exhibited over 10-fold lower TACE levels than the other periodontitis groups. TACE was positively correlated with probing pocket depth, clinical attachment levels, and RANKL concentrations in GCF. In conclusion, the increased GCF TACE levels in persons with periodontitis and their positive correlation with RANKL may indicate an association of this enzyme with alveolar bone loss, and may warrant special attention in future therapeutic approaches. [ABSTRACT FROM AUTHOR]

AIM: We investigated gingival crevicular fluid (GCF) and serum matrix metalloproteinase-8 (MMP-8) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) levels from renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), from patients receiving CsA therapy and having no GO and patients receiving tacrolimus therapy. MATERIAL AND METHODS: GCF samples were collected from sites with GO (GO+) and without GO (GO-) in CsA patients having GO; and GO- sites in CsA patients having no GO; sites from tacrolimus, gingivitis and healthy subjects. GCF and serum MMP-8 and TIMP-1 levels were determined by a time-resolved immunofluorometric assay (IFMA) and enzyme-linked immunosorbent assay. RESULTS: GO+ sites in CsA patients having GO had elevated GCF MMP-8 levels compared with those of CsA patients having no GO, tacrolimus and healthy subjects (p<0.005), but these levels were similar to those of gingivitis. The GCF MMP-8 level was higher in GO+ compared with GO- sites in CsA patients having GO (p<0.05). GCF TIMP-1 levels were similar between groups. Tacrolimus patients had lower GCF MMP-8 levels than gingivitis (p<0.005), but levels similar to the healthy group. CONCLUSION: These results show that CsA and tacrolimus therapy has no significant effect on GCF MMP-8 levels, and gingival inflammation seems to be the main reason for their elevations. [ABSTRACT FROM AUTHOR]

Background and Objective: Receptor activator of nuclear factor-κB ligand (RANKL) is responsible for the induction of osteoclastogenesis and bone resorption, whereas its decoy receptor, osteoprotegerin, can directly block this action. Because this dyad of cytokines is crucial for regulating the bone remodelling process, imbalances in their expression may cause a switch from the physiological state to enhanced bone resorption or formation. This study investigated the mRNA expression of RANKL and osteoprotegerin, as well as their relative ratio, in the gingival tissues of patients with various forms of periodontal diseases. Material and Methods: Gingival tissue was obtained from nine healthy subjects and 41 patients, who had gingivitis, chronic periodontitis, generalized aggressive periodontitis, and chronic periodontitis and were receiving immunosuppressant therapy. Quantitative real-time polymerase chain reaction was employed to evaluate the mRNA expression of RANKL and osteoprotegerin in these tissues. Results: Compared with healthy individuals, patients in all periodontitis groups, but not those with gingivitis, exhibited stronger RANKL expression and a higher relative RANKL/osteoprotegerin ratio. In addition, osteoprotegerin expression was weaker in patients with chronic periodontitis. When patients with generalized aggressive periodontitis and chronic periodontitis were compared, the former exhibited stronger RANKL expression, whereas the latter exhibited weaker osteoprotegerin expression, and there was no difference in their relative ratio. When chronic periodontitis patients were compared with chronic periodontitis patients receiving immunosuppressant therapy, osteoprotegerin, but not RANKL, expression was stronger in the latter. Conclusion: This study demonstrates that RANKL and osteoprotegerin expression are differentially regulated in various forms of periodontitis, and the relative RANKL/osteoprotegerin ratio appears to be indicative of disease occurrence. This information may confer diagnostic and therapeutic value in periodontitis. [ABSTRACT FROM AUTHOR]

By mistakes we learn: determination of matrix metalloproteinase-8 and tissue inhibitor of matrix metalloproteinase-1 in serum yields doubtful results. [ABSTRACT FROM AUTHOR]

Aim: This prospective study investigated the salivary proteome before and after periodontal therapy., Materials and Methods: Ten systemically healthy, non-smoking, stage III, grade C periodontitis patients underwent non-surgical periodontal treatment. Full-mouth periodontal parameters were measured, and saliva (n = 30) collected pre- (T0), and one (T1) and six (T6) months post-treatment. The proteome was investigated by label-free quantitative proteomics. Protein expression changes were modelled over time, with significant protein regulation considered at false discovery rate <0.05., Results: Treatment significantly reduced bleeding scores, percentages of sites with pocket depth ≥5 mm, plaque and gingival indexes. One thousand seven hundred and thirteen proteins were identified and 838 proteins (human = 757, bacterial = 81) quantified (≥2 peptides). At T1, 80 (T1 vs. T0: 60↑:20↓), and at T6, 118 human proteins (T6 vs. T0: 67↑:51↓) were regulated. The salivary proteome at T6 versus T1 remained stable. Highest protein activity post- versus pre-treatment was observed for cellular movement and inflammatory response. The small proline-rich protein 3 (T1 vs. T0: 5.4-fold↑) and lymphocyte-specific protein 1 (T6 vs. T0: 4.6-fold↓) were the top regulated human proteins. Proteins from Neisseria mucosa and Treponema socranskii (T1 vs. T0: 8.0-fold↓, 4.9-fold↓) were down-regulated., Conclusions: Periodontal treatment reduced clinical disease parameters and these changes were reflected in the salivary proteome. This underscores the potential of utilizing saliva biomarkers as prognostic tools for monitoring treatment outcomes., (© 2024 The Authors. Journal of Clinical Periodontology published by John Wiley & Sons Ltd.)

Background: In periodontitis, the equilibrium between bone formation and resorption skews in favor of bone loss. Periodontal ligament-associated protein-1 (PLAP-1) and sclerostin play a significant role in the suppression of bone formation. Tumor necrosis factor-alpha (TNF-α) is a central proinflammatory cytokine related to periodontal bone loss. This study aims to assess gingival crevicular fluid (GCF) PLAP-1, sclerostin, and TNF-α levels in individuals with periodontal disease., Methods: Seventy-one individuals diagnosed with generalized stage III grade C periodontitis (n = 23), gingivitis (n = 24), and periodontal health (n = 24) were included in the study. Full-mouth clinical periodontal measurements were performed. PLAP-1, sclerostin, and TNF-α total amounts in GCF were quantified by ELISA. Nonparametric methods were used for the data analyses., Results: Periodontitis group exhibited significantly higher GCF PLAP-1, sclerostin and TNF-α levels compared with gingivitis and periodontally healthy groups (p < 0.05). GCF PLAP-1 and TNF-α levels of gingivitis group were higher than healthy controls (p < 0.05) whereas GCF sclerostin levels were similar in two groups (p > 0.05). Significant positive correlations were found between GCF PLAP-1, sclerostin and TNF-α levels and all clinical parameters (p < 0.01)., Conclusions: To our knowledge, this is the first study showing GCF PLAP-1 levels in periodontal health and disease. Increased GCF PLAP-1 and sclerostin levels and their correlations with TNF-α in periodontitis imply that those molecules might be involved in the pathogenesis of periodontal disease. Further studies in larger mixed cohorts are needed to enlighten the possible role of PLAP-1 and sclerostin in periodontal bone loss., (© 2023 The Authors. Journal of Periodontology published by Wiley Periodicals LLC on behalf of American Academy of Periodontology.)

Objectives: This study aims to evaluate GCF Galectin-3 and Interleukin-1 beta (IL-β) levels in different grades (B and C) of stage 3 periodontitis, concurrently, and also to investigate their discriminative efficiencies in periodontal diseases., Materials and Methods: A total of 80 systemically healthy and non-smoker individuals, 20 stage 3 grade C (S3GC) periodontitis 20 stage 3 grade B (S3GB) periodontitis, 20 gingivitis, and 20 periodontally healthy were enrolled. Clinical periodontal parameters were recorded and GCF Galectin-3 and IL-1β total amounts were measured by ELISA. Receiver operating characteristics curve was used for estimating the area under the curve (AUC)., Results: Galectin-3 and IL-1β were detected in all participants. Both periodontitis groups had significantly higher GCF Galectin-3 total amounts than periodontally healthy controls (p <0.05). S3GC periodontitis group had also significantly higher GCF Galectin-3 levels than gingivitis group (p <0.05). GCF IL-1β levels in periodontitis groups were higher than gingivitis and periodontally healthy groups (p <0.05). Galectin-3 exhibited an AUC value of 0.89 with 95% sensitivity to discriminate S3GC periodontitis from periodontal health, an AUC value of 0.87 with 80% sensitivity to discriminate S3GC periodontitis versus gingivitis, while an AUC value of 0.85 with 95% sensitivity to discriminate S3GB periodontitis from healthy controls., Conclusions: GCF Galectin-3 levels are involved in the pathogenesis of periodontal diseases. Galectin-3 showed excellent diagnostic performances to discriminate S3GB and S3GC periodontitis from periodontal health and gingivitis., Clinical Relevance: The present findings suggest that GCF Galectin-3 levels may be useful in the diagnosis of the periodontal diseases., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Secretory leukocyte protease inhibitor (SLPI) is an anti-protease that protects mucosal tissue integrity owing to its anti-microbial and immunomodulatory properties. This study aimed to investigate SLPI levels in periodontal diseases, and analyze the potential correlation with clinical periodontal parameters. Whole saliva samples were obtained from healthy (n = 24), gingivitis (n = 24) and patients with stage 3 grade C periodontitis (n = 24). SLPI was measured by ELISA and normalized by total protein. Receiver operating characteristics (ROC) curve was used for estimating the area under the curve (AUC). The normalized SLPI levels were significantly reduced in periodontitis compared with gingivitis (4.84-fold) or health (1.83-fold) and negatively correlated with periodontal parameters. The ROC curves showed a good predictor value of the SLPI for differentiation of periodontitis versus health or gingivitis (AUC ≥ 0.80). This study demonstrates that the levels of SLPI are high in periodontal health, further elevated in gingivitis, but eventually decreased in severe periodontitis beyond the former two states. This observation may have broader implications in the context of inflammatory diseases affecting the oral mucosa, as it shows that the bacterial burden is disturbing the homeostatic balances of anti-microbial and anti-protease factors in the oral cavity., (© 2022. The Author(s).)

Objective: This study aimed to investigate the levels of trefoil factor family (TFF)-1, TFF-3 and interleukin (IL)-1β in gingival crevicular fluid (GCF), saliva and serum of patients with gingivitis, stage 3 periodontitis and healthy individuals., Materials and Methods: A total of 100 individuals consisting of 25 periodontally healthy, 25 gingivitis and 50 stage 3 periodontitis, were enrolled in the study. Clinical periodontal examinations were recorded and GCF, saliva and serum samples were obtained. TFF-1, TFF-3 and IL-1β were measured by ELISA., Results: TFF-1 and TFF-3 levels in both GCF, saliva and serum were higher in periodontitis patients than healthy controls (p < .001) and gingivitis group (p < .01). The levels of these peptides in all biofluids were similar between gingivitis and healthy control groups (p > .05). GCF, saliva and serum IL-1β levels were also higher in periodontitis patients than the controls (p < .01). Periodontitis patients had elevated GCF and saliva IL-β levels than gingivitis group (p < .001)., Conclusion: Elevated TFF-1 and TFF-3 levels both locally and systemically in periodontitis in parallel to increased IL-1β levels might suggest that these peptides are involved in host response during the periodontal tissue destruction., (© 2021 Wiley Periodicals LLC.)

Oral health is important not only due to the diseases emerging in the oral cavity but also due to the direct relation to systemic health. Thus, early and accurate characterization of the oral health status is of utmost importance. There are several salivary biomarkers as candidates for gingivitis and periodontitis, which are major oral health threats, affecting the gums. These need to be verified and validated for their potential use as differentiators of health, gingivitis and periodontitis status, before they are translated to chair-side for diagnostics and personalized monitoring. We aimed to measure 10 candidates using high sensitivity ELISAs in a well-controlled cohort of 127 individuals from three groups: periodontitis (60), gingivitis (31) and healthy (36). The statistical approaches included univariate statistical tests, receiver operating characteristic curves (ROC) with the corresponding Area Under the Curve (AUC) and Classification and Regression Tree (CART) analysis. The main outcomes were that the combination of multiple biomarker assays, rather than the use of single ones, can offer a predictive accuracy of > 90% for gingivitis versus health groups; and 100% for periodontitis versus health and periodontitis versus gingivitis groups. Furthermore, ratios of biomarkers MMP-8, MMP-9 and TIMP-1 were also proven to be powerful differentiating values compared to the single biomarkers.

The Objective: This study aimed to analyze active matrix metalloproteinase (aMMP-8) levels in gingival crevicular fluid (GCF), saliva and serum in the context of new criteria of gingivitis and stage 3 grade C periodontitis., The Background: Periodontal disease is an inflammatory process that can result in tooth loss and also is considered a modifying factor for systemic health. Matrix metalloproteinase (MMP)-8 is the major collagenase of periodontal tissue breakdown., Methods: Totally 83 systemically healthy and non-smoker individuals consisting of 23 periodontally healthy, 20 gingivitis and 40 stage 3 periodontitis, were recruited to the study. Clinical periodontal examinations of probing depth (PD), clinical attachment loss (CAL), gingival index (GI), plaque index (PI) and bleeding on probing (BOP) were recorded; and GCF, saliva and serum samples were obtained. aMMP-8 was measured by immunofluorometric assay (IFMA)., Results: GCF and serum aMMP-8 levels were significantly increased in periodontitis and gingivitis compared to healthy ones (P < .001), whereas gingivitis and periodontitis patients showed similar levels of aMMP-8 in GCF and serum (P > .05). Saliva levels of aMMP-8 were higher in periodontitis patients than both gingivitis and healthy individuals (P < .001). There was no significant difference in salivary aMMP-8 levels between gingivitis group and healthy controls (P > .05)., Conclusion: These findings support the involvement of aMMP-8 in periodontal diseases and suggest that its local and systemic levels can reflect stage 3 grade C periodontitis. Moreover, aMMP-8 in GCF and serum seems to have a potential to differentiate between gingivitis and periodontal health., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Hypoxia-inducible angiogenic pathway involving hypoxia inducible factor-1 alpha (HIF-1α), vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) may regulate several biological processes related to inflammation. The present study aimed to assess the effect of non-surgical periodontal treatment on gingival crevicular fluid (GCF) HIF-1α, VEGF, and TNF-α levels in generalized aggressive periodontitis (G-AgP)., Methods: Twenty G-AgP patients and 20 periodontally healthy individuals were included. G-AgP patients received scaling and root planning (SRP), per quadrant at a 1-week-interval, performed with ultrasonic and periodontal hand instruments. GCF samples were collected and clinical periodontal parameters including probing depth, clinical attachment level, gingival index and plaque index were recorded at baseline, 1 and 3 months after treatment. Biomarker levels in GCF were analyzed by ELISA., Results: At baseline all clinical parameters and GCF HIF-1α, VEGF, and TNF-α levels were significantly higher in G-AgP patients compared to healthy control (P < 0.05). All clinical parameters improved over the 3-month-period in G-AgP patients (P < 0.05). GCF HIF-1α levels in G-AgP reduced at 1 and 3 months post-treatment, however, this did not reach to statistical significance (P > 0.05). GCF VEGF and TNF-α levels remained unchanged throughout the study period (P > 0.05)., Conclusions: Within the limitations of the present study, although HIF-1α seems to possess a potential diagnostic value for G-AgP, it might not be a proper predictor of clinically favorable treatment outcome. SRP plus different adjunctive therapies could provide better information about the prognostic role of hypoxia-inducible angiogenic pathway in G-AgP., (© 2020 The Authors. Journal of Periodontology published by Wiley Periodicals, Inc. on behalf of American Academy of Periodontology.)

Background: To compare the effects of full-mouth disinfection (FMD) and full-mouth ultrasonic debridement (FMUD) on clinical, microbiological and biochemical parameters with conventional quadrant-wise scaling and root planning (Q-SRP) in severe chronic periodontitis., Methods: In the present prospective randomized controlled clinical trial with three parallel arms (#NCT04038801), 60 chronic periodontitis patients were randomly assigned to three study groups by a consecutive number in ascending order: FMD (n = 20), FMUD (n = 20), and Q-SRP (n = 20). All measurements and treatments were performed by the same investigator. At baseline, gingival crevicular fluid (GCF) and subgingival plaque were collected and clinical periodontal parameters were recorded. Ultrasonic debridement was completed within 24 hours in FMD and FMUD groups. Chlorhexidine gluconate was used for FMD. Q-SRP was performed by hand instruments per quadrant at 1-week-intervals. Clinical measurements and sampling were repeated at 1, 3, and 6 months after treatment. Real-time PCR was used for quantitative analysis of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum, and total bacteria count. GCF Calprotectin, osteocalcin, and N-telopeptide of type I collagen (NTx) levels were analyzed by ELISA. The changes of GCF biomarker levels after treatment between groups were the primary outcomes., Results: No harm was observed. All treatment strategies resulted in significant improvements in all clinical parameters (P < 0.05), with no significant differences between study groups at all time-points (P ˃ 0.05). Aggregatibacter actinomycetemcomitans was significantly decreased in FMD compared to FMUD and Q-SRP at 6 months (P < 0.05). Although GCF NTx total amounts increased in all groups during the study period, this increase was less prominent in full-mouth groups at three time points after treatment (P < 0.05)., Conclusions: Present results represent the short-term effects. Full-mouth treatment approaches offered limited beneficial effects on microbiological and biochemical parameters over quadrant-wise approach. All three treatment strategies can be recommended in the management of severe chronic periodontitis., (© 2020 American Academy of Periodontology.)

Purpose: Periodontitis is linked to a localized dysbiotic microbial shift. This trending may often not be evident due to deep taxonomic changes of low abundance organisms and lack of consideration of variations in the treatment response. By using next generation sequencing this study aims to evaluate the salivary microbiome dynamics of periodontal treatment and the implication of treatment outcome EXPERIMENTAL DESIGN: Patients with generalized aggressive periodontitis are treated non-surgically and followed up for 6 months. Saliva is collected for microbiome profiling by next generation sequencing and diversity analysis, as well as quantitative real-time polymerase chain reaction (qPCR). The treatment outcome on the first follow-up is also considered., Results: Clinical parameters are significantly improved following treatment, but with no accompanying relative abundance changes on the phylum, genus and species levels, or diversity indices. Distinctive differences are observed on species level when the sensitive qPCR is used. Patients responding poorly to treatment display a marginally lower microbiome profile distance from baseline, compared to those responding favorably., Conclusion and Clinical Relevance: Periodontal treatment does not alter the broader salivary microbiome profile, but may have selective implications on the species level. Treatment outcome can be impactful in the microbiome profile, as reduced microbiome changes may be associated with poorer clinical responses., (© 2020 The Authors. Proteomics - Clinical Applications published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Hypoxia-inducible factor-1 alpha (HIF-1α) is expressed as an adaptive response to hypoxia, mediates angiogenesis through the expression of vascular endothelial growth factor (VEGF) and can be induced by tumor necrosis factor-alpha (TNF-α). This study aimed to investigate the gingival crevicular fluid (GCF) and salivary HIF-1α, VEGF, and TNF-α levels in periodontal health and disease., Methods: A total of 87 individuals, 20 generalized aggressive periodontitis (G-AgP), 20 chronic periodontitis (CP), 26 gingivitis patients, and 21 periodontally healthy individuals, were included. Clinical periodontal parameters were recorded; GCF and salivary samples were collected; and HIF-1α, VEGF, and TNF-α levels were measured by enzyme-linked immunosorbent assay. Nonparametric tests were used for the statistical analyses., Results: G-AgP and CP groups had significantly higher GCF HIF-1α, VEGF, and TNF-α total amounts than gingivitis and healthy groups (P < 0.05). GCF HIF-1α and TNF-α total amounts in gingivitis group were significantly higher than the healthy group (P < 0.05). GCF and salivary concentrations of biomarkers were similar in both periodontitis groups (P > 0.05). Salivary HIF-1α concentrations in gingivitis group were significantly higher than G-AgP and healthy groups (P < 0.05). GCF HIF-1α, VEGF, and TNF-α total amounts were positively correlated with the site-specific clinical periodontal parameters and with each other (P < 0.05)., Conclusions: HIF-1α is detectable in GCF and saliva of periodontally diseased and healthy individuals, and the GCF levels of the biomarker can be affected by disease status. Increased GCF HIF-1α, VEGF, and TNF-α levels in both chronic and aggressive form of periodontitis might suggest the role of TNF-α/HIF-1α/VEGF pathway in the pathogenesis of periodontal diseases., (© 2018 American Academy of Periodontology.)

Objectives: To evaluate the Interleukin-4 (IL-4), bone-specific alkaline phosphatase (BALP), and C-telopeptide of type I collagen (CTX-I) levels in peri-miniscrew crevicular fluid (PMCF) during orthodontic tooth movement between 75 and 150 g of distalization force., Materials and Methods: Thirty miniscrews were placed bilaterally between the maxillary second premolars and first molars. The right and the left maxillary canines were moved distally using either 75 or 150 g of force. PMCF samples were collected before loading (T0); at 2 hours (T1) and 24 hours (T2) later; and on days 7 (T3), 14 (T4), 21 (T5), 30 (T6), and 90 (T7) after force application. Enzyme-linked immunosorbent assay kits were used to determine BALP, CTX-I, and IL-4 levels., Results: There was no significant difference between the force groups at all time points with respect to BALP, CTX-I, and IL-4 levels ( P > .05). There was no significant difference among time points for the two force groups in terms of BALP and IL-4 levels ( P > .05). The CTX-I level at T3 was significantly higher than at T0 for both force groups ( P < .05)., Conclusions: Both 75 g and 150 g of orthodontic force are within optimal force limits, and there is no difference in biochemical markers of bone turnover.

Objective: To compare effect of two different orthodontic forces on maxillary canine distalization via evaluation of 30 analytes including cytokines, growth factors, and chemokines in gingival crevicular fluid (GCF) obtained from tension and compression sites., Design: Longitudinal, split-mouth, randomized controlled trial., Methods: The upper right and left canines were randomly distalized by a continuous force of either 75 or 150 g, in 15 individuals with Class II division 1 malocclusion. GCF samples were obtained from the tension and the pressure sides of each canine at appliance placement (baseline) and after force application at 24 hours and 28 days without reactivation of the coil spring. The protein content of GCF was analysed by a multiplexed immunoassay. The effects of force, side, and time on the analyte levels were assessed by the Brunner-Langer method., Outcome: The changes of GCF analyte levels from baseline to 24 hours and 28 days., Randomization: Coin flipping was used for allocation of two forces., Blinding: The participants and periodontist who performed clinical measurements and GCF sampling were blinded to group assignment and interventions (double-blinded trial)., Results: All patients completed the study. No harm was observed. When compared to baseline, both forces caused significant up-regulation of tumour necrosis factor-α and interleukin (IL)-1RA in the tension and the pressure sides at 28 days (P < 0.05), but not at 24 hours. Although GCF volume was similar between the two force groups over time (P > 0.05), IL-8 and MCP-1 levels in GCF were significantly lower at the pressure sites receiving higher force (150 g) at 24 hours (P < 0.05)., Limitations: Although sample size (15 patients, 30 teeth) was adequate according to the initial power calculation, borderline significances may indicate lack of power or large variability among the samples., Conclusions: Although a higher force of 150 g did not result in increased cumulative canine movement or GCF production, selective host mediators were differentially regulated by the magnitude and duration of the force., Registration and Trial Protocol: The trial was registered retrospectively in the U.S. National Institutes of Health Clinical Trials Registry. Full details of trial protocol NCT03555747 are available on request., (© The Author(s) 2018. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Background: This study aimed to evaluate the levels of resistin and tumor necrosis factor-alpha (TNF-α) in gingival crevicular fluid (GCF) and saliva of obese children with gingivitis., Methods: One-hundred and thirty children (65 obese and 65 normal weight; age range 8 to 12 years) were recruited for the study. The children were classified into four subgroups based on their body mass and periodontal status; 1) obese children with gingivitis (OG, n = 33); 2) obese children with healthy periodontium (OH, n = 32); 3) normal weight children with gingivitis (NWG, n = 32); 4) normal weight children with healthy periodontium (NWH, n = 33). Body mass index (BMI) percentile, probing pocket depth (PPD), gingival index (GI), and plaque index (PI) were recorded. Resistin and TNF-α were analyzed in GCF and saliva samples by ELISA., Results: Obese children had higher BMI percentiles than normal weight children (p < 0.0001). PPD, GI, PI, GCF volume, GCF, and salivary resistin and TNF-α levels were similar between obese and normal weight children (P > 0.05). OG and NWG subgroups had significantly higher GI, PI, GCF volume, GCF resistin total amounts, and salivary resistin concentrations but lower GCF resistin and TNF-α concentrations than OH and NWH (P < 0.0001 for all). GCF resistin total amounts were positively correlated with GI, PI, and GCF TNF-α total amounts (P < 0.05)., Conclusions: To our knowledge, this is the first study evaluated the levels of resistin in GCF and saliva of children. Obesity is not associated with GCF and salivary resistin and TNF-α levels in children in the presence of gingival inflammation., (© 2018 American Academy of Periodontology.)

Periodontal diseases are among the most prevalent worldwide, but largely silent, chronic diseases. They affect the tooth-supporting tissues with multiple ramifications on life quality. Their early diagnosis is still challenging, due to lack of appropriate molecular diagnostic methods. Saliva offers a non-invasively collectable reservoir of clinically relevant biomarkers, which, if utilized efficiently, could facilitate early diagnosis and monitoring of ongoing disease. Despite several novel protein markers being recently enlisted by discovery proteomics, their routine diagnostic application is hampered by the lack of validation platforms that allow for rapid, accurate and simultaneous quantification of multiple proteins in large cohorts. Here we carried out a pipeline of two proteomic platforms; firstly, we applied open ended label-free quantitative (LFQ) proteomics for discovery in saliva ( n = 67, including individuals with health, gingivitis, and periodontitis), followed by selected-reaction monitoring (SRM)-targeted proteomics for validation in an independent cohort ( n = 82). The LFQ platform led to the discovery of 119 proteins with at least 2-fold significant difference between health and disease. The 65 proteins chosen for the subsequent SRM platform included 50 functionally related proteins derived from the significantly enriched processes of the LFQ data, 11 from literature-mining, and four house-keeping ones. Among those, 60 were reproducibly quantifiable proteins (92% success rate), represented by a total of 143 peptides. Machine-learning modeling led to a narrowed-down panel of five proteins of high predictive value for periodontal diseases with maximum area under the receiver operating curve >0.97 (higher in disease: Matrix metalloproteinase-9, Ras-related protein-1, Actin-related protein 2/3 complex subunit 5; lower in disease: Clusterin, Deleted in Malignant Brain Tumors 1). This panel enriches the pool of credible clinical biomarker candidates for diagnostic assay development. Yet, the quantum leap brought into the field of periodontal diagnostics by this study is the application of the biomarker discovery-through-verification pipeline, which can be used for validation in further cohorts., (© 2018 Bostanci et al.)

Background: Trappin-2 is a potent biologically active serine protease inhibitor with anti-inflammatory properties that has also been characterized as an "alarm anti-protease." Although the importance of trappin-2 in several chronic infections has been demonstrated, its potential involvement in periodontitis remains undefined. This study aims to investigate salivary levels of trappin-2 and interleukin (IL)-1β in periodontally healthy individuals and patients with gingivitis or generalized chronic periodontitis (CP) or aggressive periodontitis (GAgP)., Methods: Whole unstimulated saliva samples were collected from 80 systemically healthy and non-smoking individuals before full-mouth periodontal examination. Trappin-2 and IL-1β were analyzed by enzyme-linked immunosorbent assay and reported as nanograms per milligram after calibration for total protein levels., Results: Correlation analysis revealed negative association between trappin-2 and IL-1β levels. Trappin-2 also showed strong negative correlation with clinical periodontal parameters, in contrast to IL-1β, which showed positive correlation. Trappin-2 levels were significantly lower in individuals with CP and GAgP, but not gingivitis, compared with healthy individuals. Reduced salivary concentrations of trappin-2 had high sensitivity and specificity to distinguish health from periodontitis., Conclusions: Trappin-2 is abundant in the saliva of individuals with healthy periodontium in line with its role as an "anti-alarm" protease. Decreased salivary trappin-2 and increased IL-1β levels in individuals with periodontitis, compared with healthy individuals, may implicate a potential antiprotease/proinflammatory cytokine imbalance, resulting in impaired host protective capacity., (© 2018 American Academy of Periodontology.)

Objective: The aim of the present study was to investigate the effect of cyclosporine-A (CsA) medication on gingival crevicular fluid (GCF) LL-37, human neutrophil peptide (HNP)1-3 and adrenomedullin (ADM) levels., Design: CsA-treated renal transplant recipients with GO (CsA GO+) and without GO (CsA GO-), tacrolimus-medicated renal transplant recipients (n = 20/group), systemically healthy subjects with gingivitis (n = 21) and individuals free of periodontal and systemic diseases (n = 20) were included in the present study. Periodontal parameters were recorded and GCF samples were obtained from the study participants. GCF LL-37, HNP1-3 and ADM levels were analyzed by enzyme-linked immunosorbent assay., Results: GCF LL-37 total amount was higher at GO+ sites than the other study sites (p < 0.05). Total amount of GCF HNP1-3 was higher in immunosuppressive treatment groups than healthy and gingivitis groups, regardless of GO presence (p < 0.05). GCF ADM total amount was similar in all study groups. GCF volume, papillary bleeding index and hyperplastic index (p < 0.05) were significantly correlated with GCF LL-37 total amounts (p < 0.05), but not with GCF HNP1-3 and ADM total amount at GO+ sites (p > 0.05)., Conclusion: Neutrophil infiltration due to extended inflammation might have increased GCF LL-37 levels at GO+ sites and contributed to the pathogenesis of CsA-induced GO., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: Variances in fibroblasts' α2β1 integrin intensity may lead to altered adhesion to type I collagen and consequently to suppression of phagocytosis which may be one of the mechanisms for drug induced gingival overgrowth. The present study aimed to evaluate the genotype and allele frequencies of α2 integrin +807 gene in renal transplant patients with and without gingival overgrowth., Material and Methods: Seventy renal transplant patients with cyclosporine A (CsA)-induced gingival overgrowth (CsA GO+) were enrolled. Renal transplant patients without GO medicated with CsA (CsA GO-; n=79) and tacrolimus (Tac; n=52) served as controls. DNA was obtained from peripheral blood and ITGA2 +807C/T polymorphism was genotyped by polymerase chain reaction and restriction fragment length polymorphism method. Clinical parameters including probing depth and plaque, papilla bleeding and hyperplasia indexes were recorded. Chi-square, Kruskal-Wallis and Mann-Whitney tests were used in statistical analysis., Results: Clinical parameters of CsA GO+ group were significantly higher than those of the CsA GO- and Tac groups (p<0.05). ITGA2 807C/T genotype and allele frequencies of study groups were similar (p>0.05)., Conclusion: Within the limits of the present study it can be concluded that ITGA2 +807 gene polymorphism is not associated with susceptibility to CsA-induced GO., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background: The aim of this cross-sectional study is to investigate gingival crevicular fluid (GCF) osteocalcin, cross-linked N-terminal telopeptide (NTx), and calprotectin levels in cyclosporin A (CsA)-induced gingival overgrowth (GO)., Methods: Forty medicated patients with CsA including 20 with GO (CsA GO+), 10 without GO (CsA GO-), 10 with GO and chronic periodontitis (CsA CP) and 60 patients with CP alone, 20 patients with gingivitis, and 20 healthy patients were enrolled. Probing depth, clinical attachment level, plaque index, and papillary bleeding index were recorded. GCF calprotectin, osteocalcin, and NTx levels were analyzed by enzyme-linked immunosorbent assay. Parametric tests were used for statistical analysis., Results: The CsA GO+ and CP groups had significantly lower GCF osteocalcin levels and osteocalcin/NTx ratio than the healthy group, whereas GCF osteocalcin levels and osteocalcin/NTx ratio in the gingivitis group were higher than the CsA GO+, CsA GO-, CsA CP, and CP groups (P <0.05). The CP group had elevated GCF calprotectin levels compared to the other study groups (P <0.05). The CsA GO+ and CsA GO- groups also had higher GCF calprotectin levels compared to the CsA CP, gingivitis, and healthy groups (P <0.05)., Conclusions: Increased GCF calprotectin and decreased GCF osteocalcin levels in the CsA GO+ and CsA GO- groups might suggest that CsA plays a role on the levels of these markers. The similarity of GCF osteocalcin, NTx, and calprotectin levels in the CsA GO+ and CsA GO- groups might suggest that these molecules are not involved in the pathogenesis of GO.

Aim: The aim of the present study was to investigate gingival crevicular fluid (GCF) calprotectin, osteocalcin and cross-linked N-terminal telopeptide (NTx) levels in health along with different periodontal diseases., Material and Methods: Twenty chronic periodontitis (CP), 20 generalized aggressive periodontitis (G-AgP), 20 gingivitis and 20 healthy subjects were included. Probing depth, clinical attachment level, plaque index and papillary bleeding index was recorded. GCF calprotectin, osteocalcin and NTx levels were analyzed by enzyme-linked immunosorbent assay (ELISA)., Results: CP, G-AgP and gingivitis groups had higher GCF calprotectin total amount compared to healthy subjects (p< 0.008). CP and G-AgP groups had similar, but higher levels compared to gingivitis groups (p< 0.008). CP and G-AgP groups had lower GCF osteocalcin total amount compared to gingivitis and healthy groups (p< 0.008). CP group had higher GCF NTx but lower osteocalcin total amount and osteocalcin/NTx ratio than the G-AgP group (p< 0.008)., Conclusions: Our results suggest that elevated GCF calprotectin levels play a role as a reliable inflammatory marker in the pathogenesis of periodontal disease. Fluctuating GCF levels of osteocalcin and NTx might point out to the abnormal bone turnover in periodontitis. Our data document for the first time the role of NTx in the pathogenesis of different periodontal diseases.

Aim: The rationale of this study was to address whether local or systemic changes reflect proteolytic (matrix metalloproteinase-13) or oxidative (myeloperoxidase) stress in renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), in patients receiving CsA therapy and having no GO and patients receiving tacrolimus therapy., Material and Methods: Gingival crevicular fluid (GCF) samples were collected from sites with (GO+) and without GO (GO-) in CsA patients having GO; GO- sites in CsA patients having no GO; sites from tacrolimus, gingivitis and healthy subjects. GCF and serum myeloperoxidase (MPO) and matrix metalloproteinase-13 (MMP-13) levels were determined by ELISA., Results: GO+ sites in CsA patients having GO had elevated GCF MPO levels than those of CsA patients having no GO, tacrolimus and healthy subjects (p<0.005), but comparable to those of gingivitis. GCF MPO levels were higher in GO+ compared to GO- sites in CsA patients having GO (p<0.05). Patient groups had similar, but higher GCF MMP-13 levels than healthy group., Conclusions: These results show that CsA and tacrolimus therapy have not a significant effect on GCF MPO and MMP-13 levels, and gingival inflammation seems to be the main reason for their elevations., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Background: The contribution of nitric oxide (NO) to immune response and matrix degradation in the periodontal environment suggests a role for NO and NO-synthase (NOS) activity in the pathogenesis of cyclosporin A (CsA)-induced gingival overgrowth (GO). However, current knowledge on this topic is limited to experimental animal studies. The present study was undertaken on the basis of a hypothesis whether altered nitrite/nitrate levels in gingival crevicular fluid (GCF) and endothelial NOS (eNOS) and inducible NOS (iNOS) immunoreactivity in gingiva of CsA-treated patients contribute to the pathogenesis of CsA-induced GO., Methods: Twenty-four CsA-medicated renal transplant patients with GO (GO+; n = 12) or without GO (GO-; n = 12), 10 gingivitis, and 10 healthy subjects were included in the study. GCF samples from two proximal sites facing interdental papilla were collected, and papilla was excised. iNOS and eNOS were determined by immunohistochemistry. GCF nitrite/nitrate levels were analyzed based on the Griess reaction., Results: Weak iNOS immunostaining was observed in the healthy and GO- groups. In the gingivitis and GO+ groups, iNOS immunostaining significantly increased in connective tissue. Epithelial immunostaining of iNOS was localized to basal keratinocytes and the lower layer of stratum (str.) spinosum in the gingivitis group. In the GO+ group, iNOS immunostaining was differentially localized to keratinocytes of str. superficiale but considerably decreased in the str. basale. Weak eNOS immunostaining was found in the healthy and GO- groups, whereas higher immunostaining was observed in the gingivitis and GO+ groups. No intergroup differences were observed regarding nitrite/nitrate levels in GCF., Conclusion: CsA differentially upregulated iNOS, but not eNOS, in overgrown gingiva, which may play a pivotal role in the pathogenesis of CsA-induced GO.

Gibberellic acid (GA3), a plant growth regulator, is used commonly in agriculture. Its potential hazardous effects on human health, however, were relatively unexplored. Several studies demonstrated that in animals chronic GA3 consumption increased tumor formation and oxidative stress. Mast cells and Substance P (SP) play an important role in inflammation. Because chronic inflammation triggers serious conditions, including tumor formation, we examined changes in mast cell recruitment and activation as well as SP levels in skin and urinary bladder. Wistar Albino rats were treated with either a single GA3 dose or multiple GA3 doses for 30 days. Sub-chronic exposure to GA3 markedly increased mast cell recruitment and activation in both tissues. Treatment with 2mg/kg GA3 dose for 30 days decreased SP levels in skin and bladder. SP levels returned to control values in bladder and further increased in skin following 30-day treatment with the 20mg/kg GA3 dose. There was marked urothelial loss and inflammatory cell infiltration in bladder of 30-day GA3 treated groups. In skin, single GA3 doses also decreased SP levels and enhanced mast cell activation and recruitment. Since both SP and mast cell activation elicit inflammatory responses, these results demonstrate that exposure to plant growth regulators may increase inflammatory skin and bladder disease and that use of GA3 should be clearly monitored.

We aimed to identify whether abdominal fat distribution could affect the outcome in metastatic breast cancer (MBC) patients treated with aromatase inhibitors (AIs) or not. A total of 42 MBC patients treated with first line hormonal therapy were enrolled in this study. Factors associated with overall survival in the univariate analysis were age, c-erb-B2 expression intensity (+++ versus others by immunohistochemistry), and WHR, whereas only WHR retained significance in the multivariate analysis. Median overall survival figures were 472 days versus unreached for patients with a WHR of <0.92 and >or=0.92 (Log rank statistic=9.76, P=0.002). Similarly, the corresponding progression free survival figures for patients with a WHR of <0.92 and >or=0.92 were 423 versus 1004 days (Log rank statistic=6.37, P=0.012). This study suggests that WHR may serve as a potential predictive marker in MBC patients treated with AIs.

Background: Gingival crevicular fluid (GCF) levels of transforming growth factor-beta(1) (TGF-beta(1)) have been previously investigated in relation to the pathogenesis of cyclosporine-A (CsA)-induced gingival overgrowth (GO) but no clinical data are available regarding the GCF levels of TGF-beta(1) in patients treated with tacrolimus (Tac). However, as gingival inflammation is pronounced at sites of GO and this consequently may lead to an elevation in TGF-beta(1) levels the present study aimed to evaluate gingival crevicular fluid (GCF) TGF-beta(1) levels in renal transplant patients using CsA or Tac without GO., Methods: GCF TGF-beta(1) levels were investigated in 30 renal transplant patients without GO medicated with either CsA (n=15) or Tac (n=15). Sixteen gingivitis patients and 15 periodontally healthy subjects were selected as controls. Periodontal status was evaluated by measuring probing depth, plaque index and papilla bleeding index. The TGF-beta(1) levels were analysed by enzyme-linked immunosorbent assay., Results: Both CsA and Tac groups had significantly elevated GCF TGF-beta(1) total amount compared to gingivitis and healthy groups (p<0.008). GCF TGF-beta(1) total amount of CsA and Tac groups was similar (p>0.008). Gingivitis and healthy groups had also similar GCF TGF-beta(1) total amount (p>0.008)., Conclusions: Within the limits of the present data it is unlikely that TGF-beta(1) is an exclusive mediator of CsA- or Tac-induced GO. However, pathogenesis of GO is multifactorial and contribution of TGF-beta(1) to the interrelations between cytokines and growth factors with fibrogenic potential cannot be disregarded.

Platelet activation and aggregation are central processes in the pathophysiology of coronary heart disease. Mean platelet volume (MPV), a determinant of platelet activation, is a newly emerging risk marker for atherothrombosis. Rosuvastatin, a new hydrophilic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin), is approved as an adjunct to diet in patients with dyslipidemia. In this study, we evaluated the effects of rosuvastatin on the MPV levels in patients with uncontrolled primary dyslipidemia with hypolipidemic treatment. We selected 30 age, sex and body mass index matched patients with uncontrolled primary dyslipidemia and hypolipidemic diet treatment and 30 normolipidemic healthy subjects. Dyslipidemic patients were treated with 10 mg/day rosuvastatin for 12 weeks. Metabolic parameters and MPV were measured at baseline and after rosuvastatin treatment in dyslipidemic group. At baseline, the dyslipidemic group had significantly higher MPV levels than in the healthy control group (8.4 +/-1.2 fl vs. 8.1 +/-0 1.0 fl, p < 0.005). The level of MPV decreased significantly after rosuvastatin treatment from a mean of 8.4 +/- 1.2 fl to 8.1 +/- 1.3 fl, (p < 0.001). The changes in MPV levels with rosuvastatin treatment were not correlated to changes in plasma lipids (p > 0.05). In addition to its well-known hypolipidemic effect, rosuvastatin also possesses significant anti-platelet activation properties. This antiplatelet effect of rosuvastatin treatment could play a role in reducing cardiovascular complications in primary hyperlipidemic patients.

Background and Objective: Receptor activator of nuclear factor-kappaB ligand (RANKL) is responsible for the induction of osteoclastogenesis and bone resorption, whereas its decoy receptor, osteoprotegerin, can directly block this action. Because this dyad of cytokines is crucial for regulating the bone remodelling process, imbalances in their expression may cause a switch from the physiological state to enhanced bone resorption or formation. This study investigated the mRNA expression of RANKL and osteoprotegerin, as well as their relative ratio, in the gingival tissues of patients with various forms of periodontal diseases., Material and Methods: Gingival tissue was obtained from nine healthy subjects and 41 patients, who had gingivitis, chronic periodontitis, generalized aggressive periodontitis, and chronic periodontitis and were receiving immunosuppressant therapy. Quantitative real-time polymerase chain reaction was employed to evaluate the mRNA expression of RANKL and osteoprotegerin in these tissues., Results: Compared with healthy individuals, patients in all periodontitis groups, but not those with gingivitis, exhibited stronger RANKL expression and a higher relative RANKL/osteoprotegerin ratio. In addition, osteoprotegerin expression was weaker in patients with chronic periodontitis. When patients with generalized aggressive periodontitis and chronic periodontitis were compared, the former exhibited stronger RANKL expression, whereas the latter exhibited weaker osteoprotegerin expression, and there was no difference in their relative ratio. When chronic periodontitis patients were compared with chronic periodontitis patients receiving immunosuppressant therapy, osteoprotegerin, but not RANKL, expression was stronger in the latter., Conclusion: This study demonstrates that RANKL and osteoprotegerin expression are differentially regulated in various forms of periodontitis, and the relative RANKL/osteoprotegerin ratio appears to be indicative of disease occurrence. This information may confer diagnostic and therapeutic value in periodontitis.